Lesson Title:Lesson 8 Evidence for Evolution the Molecular Basis for Evolution & Phylogeny
State Standards: · Molecular evidence substantiates the anatomical evidence for evolution and provides additional detail about the sequence in which various lines of descent branched off from one another. 5F/H2
Context of Lesson:
Students will work with a National Center for Biotechnology Information (NCBI) website to learn about how similarities in amino acid sequences for similar proteins, from different organisms relate to phylogeny and evolutionary connectedness in organisms.
The pre Lab activity will take place in the Biology Classroom while the Lab activity will take place in the Computer Lab, which will be reserved in advance.
Materials: Pre-printed handouts highlighters computers for each student Overhead projector Objectives:
Students will demonstrate an understanding of a use of Bioinformatics, in the study of evolution by navigating through the NCBI website, to find connectedness between organisms based on their molecular similarities.
Students will demonstrate an understanding of phylogeny by creating a cladogram from their findings in their web-search.
Instruction: Lesson 8 Day 1 Opening:
Class time
Activity
Description
5 min
Settle in
Meet and greet students at the door of the classroom
Take role
Assume a position in front of the class and ask for attention as we begin
I will Collect any homework or class-work that is forthcoming
5 min
Opening
I will address the class “Let’s start with your 15 minutes of fun. “Does anyone have any questions regarding yesterday’s activities?”
Q&A and we are on our way.
5 min
Pairing & direction
As the class questions come to an end I will distribute the Molecular Evidence for Evolution handout.
Students will be asked to pair up with a partner of their choice as I get ready to give direction to the class
Pre Lab Activity:
Genes code for amino acids, amino acids code for proteins and proteins build body structures. Therefore, one way to observe the relatedness of species is to examine their genetic sequences. Generally, the more similarity that exists in these species, the more likely species are to be related to each other.
The amino acids below are found in the protein beta hemoglobin. This protein is generally 146 amino acids in length. To conserve space, only the amino acids where differences exist have been provided to you.
Amino Acid Number
Species 1
Species 2
Species 3
Species 4
Species 5
1
Valine
Valine
Threonine
Valine
Valine
2
Histidine
Histidine
Leucine
Histidine
Histidine
4
Threonine
Threonine
Serine
Threonine
Threonine
5
Proline
Proline
Alanine
Proline
Glycine
8
Lysine
Lysine
Aspartic Acid
Lysine
Lysine
9
Serine
Asparagine
Alanine
Serine
Serine
10
Alanine
Alanine
Histidine
Alanine
Alanine
12
Threonine
Threonine
Threonine
Threonine
Alanine
13
Alanine
Threonine
Serine
Alanine
Alanine
21
Aspartic Acid
Aspartic Acid
Glutamic Acid
Aspartic Acid
Aspartic Acid
22
Glutamic Acid
Glutamic Acid
Lysine
Glutamic Acid
Glutamic Acid
33
Valine
Leucine
Valine
Valine
Valine
50
Threonine
Serine
Serine
Threonine
Threonine
52
Aspartic Acid
Aspartic Acid
Serine
Aspartic Acid
Aspartic Acid
56
Glycine
Glycine
Serine
Glycine
Serine
69
Glycine
Glycine
Serine
Glycine
Glycine
73
Aspartic Acid
Aspartic Acid
Glutamic Acid
Aspartic Acid
Aspartic Acid
76
Alanine
Asparagine
Histidine
Alanine
Alanine
87
Threonine
Glutamine
Glutamine
Threonine
Glutamine
104
Argenine
Lysine
Lysine
Lysine
Argenine
111
Valine
Valine
Serine
Valine
Valine
112
Cysteine
Cysteine
Alanine
Cysteine
Cysteine
113
Valine
Valine
Glutamic Acid
Valine
Valine
114
Leucine
Leucine
Serine
Leucine
Leucine
115
Alanine
Alanine
Glutamic Acid
Alanine
Alanine
116
Histidine
Histidine
Leucine
Histidine
Histidine
120
Lysine
Lysine
Histidine
Lysine
Lysine
121
Glycine
Glycine
Aspartic Acid
Glycine
Glycine
122
Phenylalanine
Phenylalanine
Lysine
Phenylalanine
Phenylalanine
123
Threonine
Threonine
Serine
Threonine
Threonine
125
Proline
Glutamine
Alanine
Proline
Glutamine
126
Valine
Valine
Valine
Valine
Leucine
130
Tyrosine
Tyrosine
Phenylalanine
Tyrosine
Tyrosine
The old way of doing things was to count the differences between amino acids by hand and develop a phylogenic tree. For fun, compare the differences between species 1 and the other species and record it on the chart! Use the provided highlighter to count the number of differences that exist between species 1 and the other species. Record this information in the chart below.
Number of Differences from Species 1
Species 2
Species 3
Species 4
Species 5
This method, while accurate and effective, is rather time consuming. It also only allows us to compare organisms to each other one at a time…. Lab Activity:
The new method of doing things is to utilize the vast amount of information available on the Internet.
Both of the sites you will use today access information available on the National Center for Biotechnology Information website. National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov)
The National Center for Biotechnology Information is a website sponsored by the federal government. The goal of the site is to advance science and health by providing access to biomedical and genomic information. Pre-Lab Questions: 1. In today’s activity you will compare the gene sequences between whales and different mammals. Two of the mammal groups you will compare whales to are the orders known as artiodactyls and perissodactyls. In our analysis today, we will look at the protein known as cytochrome b. What is the function of cytochrome b?
When comparing species such as whales and other mammals, why do you think we use a protein like cytochrome b?
Go to Google or some other Internet search engine. Next to each term, identify the key characteristic of each group-artiodactyls and perissodactyls. Then, in the spaces below each group, identify two examples of artiodactyls and two examples of perissodactyls.
2. The aim of our investigation today is to compare the cytochrome b sequences among different mammal groups. In the box next to “Search,” click on the dropdown menu and select the “Protein” Database. In the box next to “for” type the phrase “cytb Mammalia” and click on the “Search” button (cytb is one of the server’s abbreviations for cytochrome b).
3. Clicking on this page takes you to the results page.
Each result lists a hyperlink to the protein as well as the scientific, Latin name of the organism from which the protein was obtained.
As you scroll through the results, you will notice that there are upwards of 2,000 pages of entries for this particular protein.
Approximately how many amino acids make up a complete, mammalian cytochrome b protein?
4. As you click through the result pages, you may notice there are proteins that are fragments and are much smaller in size than the complete proteins. You want to be careful NOT to select these organisms in your investigation. All protein samples you investigate should be the full size and listed as CYTB proteins.
5. To access each individual organism, you must click on the hyperlink for the organism.
The first organism to come up in this example is Herpestes javanicus. Clicking on the blue hyperlink tells us that this organism is commonly known as:
6. This organism page has a great deal of information. In addition to the organism’s taxonomy, it provides links to research conducted on the protein, and, important for our investigation, a copy of the protein sequence for the organism is found at the bottom of the page.
Top of page
Bottom of page
Taxonomic Information
|| ||
Protein Sequence
|| || 7. The mongoose, while an interesting subject for investigation, is not a species that we want to investigate in full detail today. The topic of our investigation, you may recall is to focus on the evolution of whales. Click “Back” in your browser to return to the mammal sequence page. Once on the page, you can narrow the search even further by searching the protein database for “Cetacea,” (the mammalian order to which whales belong). 8. The first hyperlink is for the whale, Balaenoptera musculus, commonly known as the blue whale. The information provided is organized in the exact same way as the page you viewed earlier. Our interest is in the amino acid sequence information. Although, as you saw earlier, the amino acid sequence is listed at the bottom of the page, this format will be difficult for us to deal with when we conduct our analysis. The easier format can be found by clicking on the “FASTA” link found at the top of the page.
Click on this “FASTA” link
|| 9. On the FASTA sequence page, use your mouse to select and copy the sequence. This is shown for you below.
10. After you select and copy the information, open a Microsoft Word document and paste this information into your Word document. 11. Save your Word Document and modify your sequences so that they can be interpreted. To do so, you must remove any blank spaces from the headings so that the computer does not mistake letters for amino acids. In other words….. The sequence given for the blue whale in FASTA format looks like this: >gi|5835008|ref|NP_007068.1|CYTB_10457 cytochrome b [Balaenoptera musculus] MTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHIC RDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSF WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTG IPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI LRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIV GQLASILYFLLILVLMPVTSLIENKLMKW If you tried to enter this into a sequencing program, all the letters would be interpreted as amino acids, and you would not be able to accurately compare species. To avoid this problem, you must modify the information you obtained. Thus, the blue whale should be entered as: >Blue.whale MTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHIC RDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSF WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTG IPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI LRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIV GQLASILYFLLILVLMPVTSLIENKLMKW The greater than (>) symbol indicates that what comes next is a name. This means for a two or more word named organism, you must put a period in between words in the species name. For example, if you sequence a Blue Whale, you must modify its name to read as:
>Blue.Whale
If you fail to do so, the program will begin to read the w-h-a-l-e (in whale)… as amino acids!!! 12. Try out the sequencing for yourself. Use the program to find cytochrome b sequences for the following whales, marine mammals and possible land relations of whales. Find each organism’s sequence by searching the protein database using the abbreviation of the protein (cytb) and the scientific name of the organism in quotation marks.
Enter: cytb and the “Scientific Name” of the organism in this box.
|| Be sure to double check your spelling for each scientific name, and make sure that you select a full copy of the cytochrome b protein (it should be approximately 378 amino acids in length).
Organism
Common Name
Scientific Name
Order
Relationship to Whales
Whale 1
Blue Whale
Balaenoptera musculus
Cetacea
-
Whale 2
Humpback Whale
Megaptera novaeangliae
Cetacea
-
Whale 3
Sperm Whale
Physeter catodon
Cetacea
-
Whale 4
Killer Whale
Orcinus orca
Cetacea
-
Manatee/Dugong 1
Dugong
Dugong dugon
Sirenia
Manatee/Dugong 2
Caribbean Manatee
Trichechus manatus
Sirenia
Seal
Harbor Seal
Phoca vitulina
Carnivora
Sea Lion
California Sea Lion
Zalophus californianus
Carnivora
Walrus
Walrus
Odobenus rosmarus
Carnivora
Sea Otter
Sea Otter
Enhydra lutris
Carnivora
Bear
Polar Bear
Ursus maritimus
Carnivora
Land Carnivore 1
Eurasian Wolf
Canis lupus lupus
Carnivora
Land Carnivore 2
Dog
Canis lupus familiaris
Carnivora
Artiodactyl 1
Hippopotamus
Hippopotamus amphibius
Artiodactyla
Artiodactyl 2
Cow
Bos taurus
Artiodactyla
Perissodactyl 1
Horse
Equus caballus
Perissodactyla
Perissodactyl 2
Indian Rhinoceros
Rhinoceros unicornis
Perissodactyla
Elephant 1
Asian Elephant
Elephas maximus
Proboscidea
Elephant 2
African Elephant
Loxodonta africana
Proboscidea
Fill in the first column entitled, “Relationship toWhales,” by assigning a number 1-13 for each species in terms of how closely you believe they are related to whales. (The species that you think is most closely related to whales gets a 1, the next closest relative gets a 2, the next a 3, etc.)
After modifying all of your sequences, be sure tosave your Word Document!!! Part II – Sequence Alignments
13. A feature of the NCBI webpage is the Constraint Based Multiple Alignment Tool (COBALT). COBALT serves to use the resources of NCBI to compare sequences and put them into a progressive multiple alignment. From there, a simple phylogenic tree can be constructed from this alignment.
14. The COBALT page asks you to enter protein ascensions, gis, or FASTA sequences. We have already compiled this information in the first part of this exercise. Return to your Word document that contains your cytochrome sequences. Select all of your sequences, Copy them and Paste them into the box where it indicates you should paste sequences. 15. The alignment itself takes several moments to run (more so today because we are all on the site at the same time). Once it has completed, it will show you the analysis that it ran between all species. The information provided will include information on things like number of amino acids in the protein and information about the similarities and differences that occur in the sequences. You can get the program to construct a phylogenetic tree by clicking on the “Phylogenetic Tree” hyperlink in the top right hand corner of the page.
A phylogenetic tree shows the evolutionary relationship among various species that are believed to have a common ancestor. These trees also take in evolutionary time into consideration through the distances between sequences (or the “leaves” of the tree). 16. In the space below, copy the phylogram you created in today’s class. Analysis:
How close were your predictions to your phylogram?
Did you discover anything surprising about the relationship between whales and the other groups of mammals? If so, explain.
The sequences you analyzed today were amino acid sequences. Would you expect to get the same results if you compared the DNA of these organisms instead of amino acids? Explain your response.
In three or more sentences, use what you learned today to explain how protein similarity between species can be used to determine their evolutionary relationship.
State Standards:
· Molecular evidence substantiates the anatomical evidence for evolution and provides additional detail about the sequence in which various lines of descent branched off from one another. 5F/H2
Context of Lesson:
- Students will work with a National Center for Biotechnology Information (NCBI) website to learn about how similarities in amino acid sequences for similar proteins, from different organisms relate to phylogeny and evolutionary connectedness in organisms.
- The pre Lab activity will take place in the Biology Classroom while the Lab activity will take place in the Computer Lab, which will be reserved in advance.
Materials:Pre-printed handouts highlighters computers for each student
Overhead projector
Objectives:
Instruction: Lesson 8 Day 1
Opening:
Handout to students:
Molecular Evidence for Evolution Name:Pre Lab Activity:
Genes code for amino acids, amino acids code for proteins and proteins build body structures. Therefore, one way to observe the relatedness of species is to examine their genetic sequences. Generally, the more similarity that exists in these species, the more likely species are to be related to each other.
The amino acids below are found in the protein beta hemoglobin. This protein is generally 146 amino acids in length. To conserve space, only the amino acids where differences exist have been provided to you.
This method, while accurate and effective, is rather time consuming. It also only allows us to compare organisms to each other one at a time….
Lab Activity:
The new method of doing things is to utilize the vast amount of information available on the Internet.
Both of the sites you will use today access information available on the National Center for Biotechnology Information website.
National Center for Biotechnology Information (http://www.ncbi.nlm.nih.gov)
The National Center for Biotechnology Information is a website sponsored by the federal government. The goal of the site is to advance science and health by providing access to biomedical and genomic information.
Pre-Lab Questions:
1. In today’s activity you will compare the gene sequences between whales and different mammals. Two of the mammal groups you will compare whales to are the orders known as artiodactyls and perissodactyls.
In our analysis today, we will look at the protein known as cytochrome b. What is the function of cytochrome b?
When comparing species such as whales and other mammals, why do you think we use a protein like cytochrome b?
Go to Google or some other Internet search engine. Next to each term, identify the key characteristic of each group-artiodactyls and perissodactyls. Then, in the spaces below each group, identify two examples of artiodactyls and two examples of perissodactyls.
Part I – Finding Sequences
1. Open the Internet and type the NCBI homepage into your browser.
(http://www.ncbi.nlm.nih.gov)
2. The aim of our investigation today is to compare the cytochrome b sequences among different mammal groups. In the box next to “Search,” click on the dropdown menu and select the “Protein” Database. In the box next to “for” type the phrase “cytb Mammalia” and click on the “Search” button (cytb is one of the server’s abbreviations for cytochrome b).
3. Clicking on this page takes you to the results page.
Each result lists a hyperlink to the protein as well as the scientific, Latin name of the organism from which the protein was obtained.
As you scroll through the results, you will notice that there are upwards of 2,000 pages of entries for this particular protein.
Approximately how many amino acids make up a complete, mammalian cytochrome b protein?
4. As you click through the result pages, you may notice there are proteins that are fragments and are much smaller in size than the complete proteins. You want to be careful NOT to select these organisms in your investigation. All protein samples you investigate should be the full size and listed as CYTB proteins.
5. To access each individual organism, you must click on the hyperlink for the organism.
The first organism to come up in this example is Herpestes javanicus. Clicking on the blue hyperlink tells us that this organism is commonly known as:
6. This organism page has a great deal of information. In addition to the organism’s taxonomy, it provides links to research conducted on the protein, and, important for our investigation, a copy of the protein sequence for the organism is found at the bottom of the page.
7. The mongoose, while an interesting subject for investigation, is not a species that we want to investigate in full detail today. The topic of our investigation, you may recall is to focus on the evolution of whales. Click “Back” in your browser to return to the mammal sequence page.
Once on the page, you can narrow the search even further by searching the protein database for “Cetacea,” (the mammalian order to which whales belong).
8. The first hyperlink is for the whale, Balaenoptera musculus, commonly known as the blue whale.
The information provided is organized in the exact same way as the page you viewed earlier. Our interest is in the amino acid sequence information. Although, as you saw earlier, the amino acid sequence is listed at the bottom of the page, this format will be difficult for us to deal with when we conduct our analysis. The easier format can be found by clicking on the “FASTA” link found at the top of the page.
9. On the FASTA sequence page, use your mouse to select and copy the sequence. This is shown for you below.
10. After you select and copy the information, open a Microsoft Word document and paste this information into your Word document.
11. Save your Word Document and modify your sequences so that they can be interpreted. To do so, you must remove any blank spaces from the headings so that the computer does not mistake letters for amino acids. In other words…..
The sequence given for the blue whale in FASTA format looks like this:
>gi|5835008|ref|NP_007068.1|CYTB_10457 cytochrome b [Balaenoptera musculus]
MTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHIC
RDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSF
WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTG
IPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI
LRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIV
GQLASILYFLLILVLMPVTSLIENKLMKW
If you tried to enter this into a sequencing program, all the letters would be interpreted as amino acids, and you would not be able to accurately compare species. To avoid this problem, you must modify the information you obtained. Thus, the blue whale should be entered as:
>Blue.whale
MTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHIC
RDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSF
WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTG
IPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI
LRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIV
GQLASILYFLLILVLMPVTSLIENKLMKW
The greater than (>) symbol indicates that what comes next is a name. This means for a two or more word named organism, you must put a period in between words in the species name. For example, if you sequence a Blue Whale, you must modify its name to read as:
>Blue.Whale
If you fail to do so, the program will begin to read the w-h-a-l-e (in whale)… as amino acids!!!
12. Try out the sequencing for yourself. Use the program to find cytochrome b sequences for the following whales, marine mammals and possible land relations of whales. Find each organism’s sequence by searching the protein database using the abbreviation of the protein (cytb) and the scientific name of the organism in quotation marks.
Be sure to double check your spelling for each scientific name, and make sure that you select a full copy of the cytochrome b protein (it should be approximately 378 amino acids in length).
After modifying all of your sequences, be sure to save your Word Document!!!
Part II – Sequence Alignments
13. A feature of the NCBI webpage is the Constraint Based Multiple Alignment Tool (COBALT). COBALT serves to use the resources of NCBI to compare sequences and put them into a progressive multiple alignment. From there, a simple phylogenic tree can be constructed from this alignment.
http://www.ncbi.nlm.nih.gov/tools/cobalt/
14. The COBALT page asks you to enter protein ascensions, gis, or FASTA sequences. We have already compiled this information in the first part of this exercise.
Return to your Word document that contains your cytochrome sequences. Select all of your sequences, Copy them and Paste them into the box where it indicates you should paste sequences.
15. The alignment itself takes several moments to run (more so today because we are all on the site at the same time). Once it has completed, it will show you the analysis that it ran between all species. The information provided will include information on things like number of amino acids in the protein and information about the similarities and differences that occur in the sequences.
You can get the program to construct a phylogenetic tree by clicking on the “Phylogenetic Tree” hyperlink in the top right hand corner of the page.
A phylogenetic tree shows the evolutionary relationship among various species that are believed to have a common ancestor. These trees also take in evolutionary time into consideration through the distances between sequences (or the “leaves” of the tree).
16. In the space below, copy the phylogram you created in today’s class.
Analysis:
How close were your predictions to your phylogram?
Did you discover anything surprising about the relationship between whales and the other groups of mammals? If so, explain.
The sequences you analyzed today were amino acid sequences. Would you expect to get the same results if you compared the DNA of these organisms instead of amino acids? Explain your response.
In three or more sentences, use what you learned today to explain how protein similarity between species can be used to determine their evolutionary relationship.
__
Sequences
>Blue.Whale
MTNIRKTHPLMKIINDAFIDLPTPSNISSWWNFGSLLGLCLIVQILTGLFLAMHYTPDTMTAFSSVTHIC
RDVNYGWVIRYLHANGASMFFICLYAHMGRGLYYGSHAFRETWNIGVILLFTVMATAFVGYVLPWGQMSF
WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIIMALAIVHLIFLHETGSNNPTG
IPSDMDKIPFHPYYTIKDILGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI
LRSIPNKLGGVLALLLSILVLALIPMLHTSKQRSMMFRPFSQFLFWVLVADLLTLTWIGGQPVEHPYVIV
GQLASILYFLLILVLMPVTSLIENKLMKW
>Humpback.Whale
MTNIRKTHPLMKIINDTFIDLPTPSNISSWWNFGSLLGLCLIMQILTGLFLAMHYTPDTTTAFSSVTHIC
RDVNYGWIIRYLHANGASMFFICLYAHMGRGLYYGSYAFRETWNIGVILLFTVMATAFVGYVLPWGQMSF
WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIITALAIVHLIFLHETGSNNPTG
IPSNMDKIPFHPYYTIKDTLGALLLILTLLMLTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI
LRSIPNKLGGVLALLLSILILAFIPMLHTSKQRSMMFRPFSQFLFWMLVADLLTLTWIGGQPVEHPYMIV
GQLASILYFLLILVLMPMTSLIENKLMKW
>Sperm.Whale
MTNIRKSHPLMKIINNAFIDLPTPSNISSWWNFGSLLGLCLIMQILTGLFLAMHYTPDTTTAFSSITHIC
RDVNYGWTIRYLHANGASMFFICLYTHMGRGWYYGSYIFQETWNVGMMLLITVMATAFVGYVLPWGQMSF
WAATVITNLLSAIPYIGTTLVEWVWGGFSVDKATLTRFFTLHFILPFITLTLTMVHLLFLHETGSNNPTG
IPSNMDKIPFHPYHTIKDTMGALLLILSLLTLTLFAPDLLGDPDNYTPANPLNTPTHIKPEWYFLFAYAI
LRSVPNKLGGVLALLLSILILVFIPMLHTAKQRSMMFRPFSQFLFWTLIMDLLTLTWIGGQPVEHPYVTV
GQLASILYFLLILILMPTASLIENKLLKW
>Killer.Whale
MTNIRKTHPLMKILNNAFIDLPTPSNISSWWNFGSLLGLCLITQILTGLLLAMHYTPDTSTAFSSVAHIC
RDVNYGWFIRYLHANGASMFFICLYAHIGRSLYYGSYMFQETWNVGVLLLLAVMATAFVGYVLPWGQMSF
WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIITALAAVHLLFLHETGSNNPTG
IPSNMDMIPFHPYHTIKDTLGALLLILTLLALTLFAPDLLGDPDNYTPANPLSTPAHIKPEWYFLFAYAI
LRSVPNKLGGVLALLLSILILIFIPMLQTSKQRSMMFRPFSQLLFWTLIADLLTLTWIGGQPVEHPYIIV
GQLASILYFLLILVLMPTISLIENKLLKW
>Dugong
MTNIRKSHPLIKILNNSFIDLPTPVNISSWWNFGSLLGACLIIQILTGLFLAMHYTSDTLTAFSSVTHIC
RDVNYGWIIRYLHANGASMFFLCLYAHIGRGIYYGSYLYPETWNIGIVLLLTVMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTNLVEWVWGGFSVDKATLTRFFALHFILPFIVTALVMVHLLFLHETGSNNPTG
LISDSDKIPFHPYYSVKDLLGLFLLILVLLLLTLFSPDMLGDPDNYTPANPLNTPPHIKPEWYFLFRYAI
LRSIPNKLGGVLALVLSILILALLPLLHTSKQRSLSFRPLSQCLFWILVADLITLTWIGGQPVEHPYIII
GQLASILYFSIILIFMPIAGLIENHLLKW
>Caribbean.Manatee
MTNIRKSHPLIKILNDSFIDLPTPTNISSWWNFGSLLGACLIIQILTGLFLAMHYTSDTLTAFSSVAHIC
RDVNYGWIIRYLHANGASMFFLCLYAHIGRGIYYGSYLYSETWNIGIILLLTIMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTNLVEWVWGGFSVDKATLTRFFALHFTLPFVVTALVTVHLLFLHETGSNNPTG
LISDSDKIPFHPYYSIKDLLGLFLLILILLLPTLFSPDMLGDPDNYTPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALVLSILILALLPLLHTSKQRSLLFRPLSQCLFWILVTDLITLTWIGGQPVEHPYIVI
GQLASILYFSIILIFMPIAGLIENHLLKW
>Harbor.Seal
MTNIRKTHPLMKIINNSFIDLPTPSNISAWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVTHIC
RDVNYGWIIRYLHANGASMFFICLYMHVGRGLYYGSYTFTETWNIGIILLFTVMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYVGTDLVQWIWGGFSVDKATLTRFFAFHFILPFVVLALDAVHLLFLHETGSNNPSG
IMSDSDKIPFHPYYTIKDILGALLLILVLTLLVLFSPDLLGDPDNYIPPNPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALVLSILVLAIMPLLHTSKQRGMMFRPISQCLFWFLVADLLTLTWIGGQPVEHPYITV
GQLASILYFTILLVLMPIASIIENNILKW
>California.Sea.Lion
MTNIRKVHPLAKIINSSLIDLPTPSNISAWWNFGSLLAACLALQILTGLFLAMHYTSDTTTAFSSVTHIC
RDVNYGWIIRYMHANGASMFFICLYMHVGRGLYYGSYTLTETWNIGIILLFTIMATAFMGYVLPWGQMSF
WGATVITNLLSAVPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFMASALVMVHLLFLHETGSNNPSG
ISSDSDKIPFHPYYTIKDILGTLLLILTLMLLVMFSPDLLGDPDNYIPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALLLSILILAIIPLLHTSKQRGMMFRPISQCLFWLLVADLLTLTWIGGQPVEHPFITI
GQLASILYFTILLVFMPIAGIIENNILKW
>Walrus
MTNIRKTHPLAKIINNTFIDLPTPSNISAWWNFGSLLATCLILQILTGLFLAMHYTSDTTTAFSSITHIC
RDVNYGWIIRYMHANGASMFFICLYAHMGRGIYYGSYTLAETWNIGIVLLLTIMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYVGTDLVEWVWGGFSVDKATLTRFLALHFVLPFMALALTAVHLLFLHETGSNNPSG
ILSDSDKIPFHPYYTIKDILGLIILILILMLLVLFSPDLLGDPDNYTPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALLLSILVLAIVPSLHTSKQRSMMFRPISQCLFWLLVADLITLTWIGGQPVEHPFIII
GQLASILYFMILLVFMPIAGMIENSILKW
>Sea.Otter
MTNIRKTHPLTKIINNSFIDLPAPSNISAWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVAHIC
RDVNYGWIIRYMHANGASMFFICLFLHVGRGLYYGSYMFSETWNIGIVLLFTVMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFIISALAMIHLLFLHETGSNNPSG
IPSNSDKIPFHPYYTIKDILGALSLVLALTTLVLFSPDLLGDPDNYIPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALILSILILAIIPLLHTSKQRSMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFVVI
GQLASILYFMILLVLMPITSIIENNLLKW
>Polar.Bear
MTNIRKTHPLAKIINNSFIDLPTPSNISAWWNFGSLLGVCLILQILTGLFLAMHYTSDTTTAFSSVTHIC
RDVHYGWVIRYVHANGASMFFICLFMHVGRGLYYGSYLFSETWNIGIILLFTVMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFIILALAAVHLLFLHETGSNNPSG
IPSDSDKIPFHPYYTIKDILGALLLTLALATLVLFSPDLLGDPDNYIPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALIFSILILALIPLLHTSKQRGMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFIII
GQLASILYFTILLVLMPIAGIIENNLLKW
>Eurasian.Wolf
MTNIRKTHPLAKIVNNSFIDLPAPSNISAWWNFGSLLGVCLILQILTGLFLAMHYTSDTATAFSSVTHIC
RDVNYGWIIRYMHANGASMFFICLFLHVGRGLYYGSYVFMETWNIGIVLLFATMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFIIAALAMVHLLFLHETGSNNPSG
ITSDSDKIPFHPYYTIKDILGALLLLLILMSLVLFSPDLLGDPDNYTPANPLNTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALVFSILILAFIPLLHTSKQRSMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFIII
GQVASMLYFTILLILMPTVSVIENNLLKW
>Dog
MTNIRKTHPLAKIVNNSFIDLPAPSNISAWWNFGSLLGVCLILQILTGLFLAMHYTSDTATAFSSVTHIC
RDVNYGWIIRYMHANGASMFFICLFLHVGRGLYYGSYVFMETWNIGIVLLFATMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFIIAALAMVHLLFLHETGSNNPSG
ITSDSDKIPFHPYYTIKDILGALLLLLILMSLVLFSPDLLGDPDNYTPANPLNTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALVFSILILAFIPLLHTSKQRSMMFRPLSQCLFWLLVADLLTLTWIGGQPVEHPFIII
GQVASILYFTILLILMPTVSVIENNLLKW
>Hippopotamus
MTNIRKSHPLMKIINDAFVDLPAPSNISSWWNFGSLLGVCLILQILTGLFLAMHYTPDTLTAFSSVTHIC
RDVNYGWVIRYMHANGASIFFICLFTHVGRGLYYGSHTFLETWNIGVILLLTTMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTDLVEWIWGGFSVDKATLTRFFAFHFILPFVITALAIVHLLFLHETGSNNPTG
IPSNADKIPFHPYYTIKDILGILLLMTTLLTLTLFAPDLLGDPDNYTPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALALSILILALIPMLHTSKQRSLMFRPLSQCLFWALIADLLTLTWIGGQPVEHPFIII
GQVASILYFLLILVLMPVAGIIENKLLKW
>Cow
MTNIRKSHPLMKIVNNAFIDLPAPSNISSWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVTHIC
RDVNYGWIIRYMHANGASMFFICLYMHVGRGLYYGSYTFLETWNIGVILLLTVMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFIIMAIAMVHLLFLHETGSNNPTG
ISSDVDKIPFHPYYTIKDILGALLLILALMLLVLFAPDLLGDPDNYTPANPLNTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALAFSILILALIPLLHTSKQRSMMFRPLSQCLFWALVADLLTLTWIGGQPVEHPYITI
GQLASILYFLLILVLMPTAGTIENKLLKW
>Horse
MTNIRKSHPLIKIINHSFIDLPAPSNISSWWNFGSLLGICLILQILTGLFLAMHYTSDTTTAFSSVTHIC
RDVNYGWIIRYLHANGASMFFICLFIHVGRGLYYGSYTFLETWNIGIILLFTVMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTTLVEWIWGGFSVDKATLTRFFAFHFILPFIITALVVVHLLFLHETGSNNPSG
IPSDMDKIPFHPYYTIKDILGLLLLILLLLTLVLFSPDLLGDPDNYTPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALILSILILALIPTLHMSKQRSMMFRPLSQCVFWLLVADLLTLTWIGGQPVEHPYVII
GQLASILYFSLILIFMPLASTIENNLLKW
>Rhinoceros
MTNIRKSHPLVKIINHSFIDLPTPSNISSWWNFGSLLGICLILQILTGLFLAMHYTPDTTTAFSSVTHIC
RDVNYGWMIRYLHANGASMFFICLFIHVGRGLYYGSYTFLETWNIGIILLFTLMATAFMGYVLPWGQMSF
WGATVITNLLSAIPYIGTNLVEWIWGGFSVDKATLTRFFAFHFILPFIILALAITHLLFLHETGSNNPSG
IPSNMDKIPFHPYYTIKDILGALLLILVLLILVLFFPDILGDPDNYTPANPLSTPPHIKPEWYFLFAYAI
LRSIPNKLGGVLALAFSILILLLIPYLHTSKQRSMMFRPLSQCMFWLLVADLLTLTWIGGQPVEHPFIII
GQLASILYFSLILVLMPLAGIIENNLLKW
>Indian.Elephant
MTHTRKSHPLFKIINKSFIDLPTPSNISTWWNFGSLLGACLITQILTGLFLAMHYTPDTMTAFSSMSHIC
RDVNYGWIIRQLHSNGASIFFLCLYTHIGRNIYYGSYLYSETWNTGIMLLLITMATAFMGYVLPWGQMSF
WGATVITNLFSAIPYIGTNLVEWIWGGFSVDKATLNRFFAFHFILPFTMVALAGVHLTFLHETGSNNPLG
LTSNSDKIPFHPYYTIKDFLGLLILILLLLLLALLSPDMLGDPDNYMPADPLNTPLHIKPEWYFLFAYAI
LRSVPNKLGGVLALFLSILILGLMPFLHTSKHRSMMLRPLSQVLFWTLTMDLLTLTWIGSQPVEYPYTII
GQMASILYFSIILAFLPIAGMIENYLIK
>African.Elephant
MTHIRKSHPLLKIINKSFIDLPTPSNISTWWNFGSLLGACLITQILTGLFLAMHYTPDTMTAFSSMSHIC
RDVNYGWIIRQLHSNGASIFFLCLYTHIGRNIYYGSYLYSETWNTGIMLLLITMATAFMGYVLPWGQMSF
WGATVITNLFSAIPYIGTNLVEWIWGGFSVDKATLNRFFALHFILPFTMIALAGVHLTFLHETGSNNPLG
LTSDSDKIPFHPYYTIKDFLGLLILILLLLLLALLSPDMLGDPDNYMPADPLNTPLHIKPEWYFLFAYAI
LRSVPNKLGGVLALLLSILILGLMPLLHTSKHRSMMLRPLSQVLFWTLTMDLLTLTWIGSQPVEYPYIII
GQMASILYFSIILAFLPIAGVIENYLIK